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AZ St Jan Brugge Oostende, campus Oostende

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Abstract Bachelor Project MLT 2018-2019: Development of a procedure to detect CPE in a routine laboratory by comparing rapid tests and hydrolysis tests
Carbapenemase producing Enterobacterales (CPE) are bacteria that pose a danger to the public health of people all around the world. There has been a rise of CPE in Europe since it was imported by countries in the Middle East and South Asia. The purpose of this bachelor thesis is to set up a fast and accurate CPE-detection procedure for a routine laboratory to prevent spread of the CPE. The procedure will be achieved by evaluating four different kits to detect CPE: Resist-4 O.K.N.V. (Coris), β CARBATM test (Bio-Rad), Rapidec® carba NP (BioMérieux) and Rapid CARB Blue kit (Rosco). The Resist-4 O.K.N.V. of Coris is the most user friendly kit. It has a short turnaround time and is easy interpretable. The hydrolysis kit of Bio-Rad, the β CARBATM test, is better compared to the Rapidec® carba NP (BioMérieux) and Rapid CARB Blue kit (Rosco). The results of the kits showed a sensitivity varying between 87 % and 93 % and the specificity of the different kits is consistently 93%. Although the β CARBATM test (Bio-Rad) shows a lesser sensitivity, it was the only kit that was able to detect OXA-58. The CPE-detection procedure is established with the rapid test Resist-4 O.K.N.V. of CORIS and the hydrolysis test of β CARBATM test of Bio-Rad.
 
Abstract bachelorproef 2016-2017Comparison of MRSA selects II (Biorad) and Brilliance MRSA 2 (Oxoid) screening agar plates

MRSA is a major cause of infections. Therefore, a MRSA screening is performed daily at the hospital.

MRSA screening agar plates are used to detect MRSA. The purpose of this study is to compare the MRSA select II (Biorad) and Brilliance MRSA 2 (Oxoid) screening agar plates based on color colonies, the interference of the wiper fluid on the plates and the detection limit, specificity, sensitivity, PPW and NPW are determined.

The chromogenic agar are used to detect the presence of MRSA. In suspect colonies, further identification with the MALDI-TOF MS, VITEK 2 Compact, PBP2a Culture Colony test and / or cefoxitin E-test is performed.

All MRSA samples were positive on both plates. All MSSA samples were negative at MRSA select II (Biorad). Eighteen of the Twenty were negative at Brilliance MRSA 2 (Oxoid).
On the other two plates  suspicious colonies were visible. Further identification was performed and both were identified as MSSA. In the CNS samples, both plates were negative. This is as expected. At the wipers, more false positive results were observed with Brilliance MRSA 2 (Oxoid) than in MRSA select II (Biorad). Also, there were significantly more plates of Brilliance MRSA 2 (Oxoid) that were affected by the wiper. The detection limit is very good at Brilliance MRSA 2 (Oxoid) because, according to this research, it is better than what the company had specified. The sensitivity and negative predictive value were 100% for both MRSA screening agar plates and therefore very good. The specificity and positive predictive value were better at MRSA select II (Biorad). This may be due to lack of experience of interpreting the plates.

MRSA select II (Biorad) is currently being used. Due to the lack of experience, the Brilliance MRSA 2 (Oxoid) plates are less interpretable. The laboratory of the hospital AZ-Sint-Jan-Bruges-Ostend AV campus Henri Serruys is strongly considering using the Brilliance MRSA 2 (Oxoid) plate as  the plate is cheaper and to improve uniformity in the fusion hospital.

Abstract bachelorproef 2015-2016Validation of the assay of anti-TPO and anti-TG on the Cobas e411 and the VIDAS 

The purpose of this study is evaluating Cobas e411 and VIDAS in order to be able to use the best device in the laboratory. These devices are able to measure two parameters of thyroid autoantibodies, particularly anti-thyroid peroxidase (anti-TPO) and anti-thyroglobulin (anti-TG).

In some people, there are antibodies present in the blood which are directed against the thyroid. This is done by a mistake of the immune system that will attack its own cells. It sees these cells as enemy invaders. The determination of thyroid antibodies has a place in the differential diagnosis of thyroid disorders.

The Cobas e411 uses the technology of electrochemiluminescence immunoassay (ECLIA). The test principle of anti-TPO and anti-TG is based on a competition reaction. The VIDAS uses the technology of an enzyme linked fluorescent assay, based on the assay principle of a two-step sandwich ELISA technique for both anti-TPO and anti-TG.

The two devices are compared by checking the repeatability and reproducibility. A method comparison is done between the Cobas e411 and the Cobas e602 of campus Sint-Jan by using a Passing-Bablok regression analysis. Because of different reference ranges, the method comparison of the VIDAS and Cobas e602 is based on the agreement of results.

The results of the repeatability and reproducibility of anti-TPO on the Cobas e411 and the VIDAS are acceptable. The repeatability results of anti-TG on the Cobas e411 are acceptable, the VIDAS results are less acceptable only for the negative control. The reproducibility results of anti-TG on the Cobas e411 are not acceptable, the results of the VIDAS are only for the negative control LTA not acceptable.

The results of method comparison of the Cobas e411 and the Cobas e602 are acceptable. The results of method agreement between the Cobas e602 and VIDAS show discordant cases for anti-TG, where the VIDAS gives the right results.

Of cause of the results of this study and the low request for both tests, the laboratory of campus Henri Serruys gives the preference to analyse anti-TPO and anti-TG on the platform of VIDAS.

 
Samenvatting eindwerk 2014-2015: Evaluation of EBV VCA IgM and EBNA IgG on Vidas and VirClia
The purpose of this study is evaluating Vidas and VirClia in order to be able to use the best device in the laboratory. These devices are able to measure two parameters of Epstein-Barr virus.
Epstein-Barr virus is a member of the herpesvirus family and one of the most common human viruses. The virus is present worldwide, and most people become infected with EBV at some point during their lives. In general terms, antibody prevalence rates reach 95% or higher among elderly individuals in Western societies. Infectious mononucleosis is the most common disease caused by EBV, leading to fever, cervical adenopathies, splenomegaly, and pharyngitis. EBV is also involved in the origin of proliferative syndromes in immunosuppressed patients and EBV infection is as well associated with Burkitt’s lymphoma and nasopharyngeal carcinoma.
Antibodies to several antigen complexes may be measured for detection of Epstein-Bar virus. These antigens are the viral capsid antigen, the early antigen and the EBV nuclear antigen. The presence of IgM antibodies to VCA and absence of antibodie to EBNA, are indicative of primary EBV infection.
In this study only EBV VCA IgM and EBNA IgG were measured.
The assay principle of the Vidas combines an enzyme immunoassay method by immunocapture with a final fluorescent detection (ELFA). The principle of the VirClia is based on the indirect chemiluminescent immunoassa (CLIA).
Both devices were tested with an interrun and intrarun to check the accuracy. The results of the samples were compared with methods that were used on the Evolis. The sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) are calculated.
The results show five discordant results for the Vidas and only one for the VirClia. Consequently, the VirClia shows better performances in the sensitivity, specificity, PPV and NPV.
The conclusion of this study is that the VirClia obtained the best results.
 
Samenvatting eindwerk 2013-2014: Validation of the IVD MALDI Biotyper system of the firm Bruker in the daily routine of a clinical microbiological laboratory
In order to use the new MALDI-TOF routinely in the microbiological laboratory of AZ Sint-Jan campus Henri Serruys a validation of the MALDI-TOF had to be performed.
Matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry can examine the profile of proteins detected directly from an intact bacterial cell surface. This technique, which is based on relative molecular masses, is a soft-ionization method that allows desorption of peptides and proteins from different microorganisms. Ions are separated and detected according to their molecular mass and charge. Bacteria are identified on the basis of the mass/charge ratio (m/z). This approach yields reproducible spectrum. These spectra can be compared with a database and are linked to the correct microorganism. For a good validation there is need of a good comparative study. One of the most important tools used for comparison is the Vitek 2 Compact. There are also the Mini API, some biochemical tests and selective agars.
A validation plan was worked out at the hospital laboratory containing the following criteria:
  1. Accuracy – correlation study
  2. Reproducibility
  3. Controls: operator-targets-contamination
  4. Impact of different kinds of culture media
  5. Age of the cultures
  6. Validation of the toothpicks
In all of the criteria  the MALDI-TOF scored high. It may be decided that it is a good device for routine identification of bacteria and yeasts in a microbiological lab. But there are still some misidentifications mainly for the S. pneumonia/mitis/oralis group noticed in our validation. In the near future a score list system will be implemented by Bruker in order to solve this problem.
Thus, the Maldi Biotyper of the firm Bruker is an excellent method for identifying micro-organisms. It has few limitations and is easily implementable in a routine microbiology lab. The advantages are low cost, speed of execution, reliability and precision. 

Address

Koopvaardijstraat 29
8400 Oostende
059/555125
Belgium

Contacts

Traineeship supervisor
Dr. S. Van Erum
Traineeship supervisor
Kathy Boydens
058/555020
kathy.boydens@henriserruysav.be
Traineeship supervisor
Dr. Evelyne Vanderstraeten
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